Tertiary amino lipidated cationic peptides for nucleic acid delivery

Abstract

The present disclosure relates to tertiary amino lipidated and/or PEGylated cationic peptide compounds and complexes thereof with nucleic acids for endocellular delivery, methods for preparing the compounds and complexes, and methods for delivering polyanionic compounds to cells.

1 . A tertiary amino lipidated and/or PEGylated cationic peptide compound of formula (I) or a salt thereof: 
       
         
           
           
               
               
           
         
         wherein: 
         m is an integer from 0 to 10; 
         n is an integer from 0 to 5; 
         s is an integer from 0 to 5; 
         t is an integer from 0 to 10;
 wherein at least one of m, n, s, and t is nonzero; 
 
         r is an integer from 1 to 20; 
         each o is independently an integer 0, 1, or 2; 
         each q is independently an integer 0, 1, or 2; 
         each p is independently an integer 1 or 2; 
         R 1  is —H, alkyl, alkylaryl, —COR 1a  or a lipid moiety,
 wherein Ria is —H, —OH, alkyl, aryl, alkylaryl, —O-alkyl, or —O-alkylaryl; 
 
         each R 2  is independently an ethylene glycol moiety of the formula —CH 2 CH 2 O(CH 2 CH 2 O) u CH 3 , and wherein each u is independently an integer from 2 to 200; 
         each R 3  is independently a lipid moiety; 
         each R 4  is independently a neutral spacer moiety or a lipid moiety; 
         each R 5  is independently a cationic moiety; 
         each R 6  is independently an ethylene glycol moiety of the formula —CH 2 CH 2 O(CH 2 CH 2 O) v CH 3 , and wherein each v is independently an integer from 2 to 200; 
         R 7  is —H, alkyl, acyl, —OH, —OR 7a , —NH 2 , —NHR 7a , or a lipid moiety, wherein R 7a  is alkyl, acyl, or a lipid moiety; and 
         each R a  and R b  is independently —H, C 1 -C 4 -alkyl, or a side chain moiety found on a naturally- or non-naturally-occurring amino acid. 
       
     
     
         2 .- 36 . (canceled) 
     
     
         37 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 1 , wherein m is 0; n is 0; s is an integer from 1 to 5; t is 0; r is an integer from 1 to 5; o is 0; q is 2; and p is 1, and having a structure: 
       
         
           
           
               
               
           
         
       
     
     
         38 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 37 , wherein:
 R 1  is H;   each R 3  independently is C 8 -C 24 alkyl;   each R 4  is independently a neutral spacer moiety that is a C 1 -C 4 -alkyl substituted by cycloalkyl, heterocyclylalkyl, alkylaryl, arylalkyl, alkylheteroaryl, heteroarylalkyl, alkoxy, alkoxyalkyl, or hydroxyalkyl, wherein each cycloalkyl, heterocyclylalkyl, alkylaryl, arylalkyl, alkylheteroaryl, heteroarylalkyl, alkoxy, alkoxyalkyl, or hydroxyalkyl is optionally substituted with one or more substituents —OH, halo, or alkoxy;   each R 5  is independently selected from the group consisting of aminoalkyl, alkylaminoalkyl, aminoalkylaminoakyl, guanidinoalkyl, N-heterocyclylalkyl, and N-heteroaryl;   R 7  is NH 2 ; and   each of R a  and R b  is H.   
     
     
         39 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 37 , wherein
 each R 4  independently is selected from the group consisting of   
       
         
           
           
               
               
           
         
       
       and
 each R 5  is independently selected from the group consisting of 
 
       
         
           
           
               
               
           
         
       
     
     
         40 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 1 , wherein m is 0; n is 0; s is an integer from 1 to 5; t is 0; r is an integer from 2 to 10; o is 0; q is 0; and p is 1, and having a structure: 
       
         
           
           
               
               
           
         
       
     
     
         41 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 40 , wherein
 R 1  is H;   each R 3  independently is C 8 -C 24 alkyl;   each R 5  is independently selected from the group consisting of aminoalkyl, alkylaminoalkyl, aminoalkylaminoakyl, guanidinoalkyl, N-heterocyclylalkyl, and N-heteroaryl;   R 7  is NH 2 ; and   each of R a  and R b  is H.   
     
     
         42 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 40 , wherein
 each R 3  independently is   
       
         
           
           
               
               
           
         
       
       and
 each R 5  is independently selected from the group consisting of 
 
       
         
           
           
               
               
           
         
       
     
     
         43 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 1 , wherein m is 0; n is 0; s is 3 or 4; t is 0; r is 1; o is 0; q is 2; and p is 2, and having a structure: 
       
         
           
           
               
               
           
         
       
     
     
         44 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 43 , wherein:
 R 1  is H or alkyl;   each R 3  independently is C 8 -C 24 alkyl;   each R 4  is independently a neutral spacer moiety that is a C 1 -C 4 -alkyl substituted by cycloalkyl, heterocyclylalkyl, alkylaryl, arylalkyl, alkylheteroaryl, heteroarylalkyl, alkoxy, alkoxyalkyl, or hydroxyalkyl, wherein each cycloalkyl, heterocyclylalkyl, alkylaryl, arylalkyl, alkylheteroaryl, heteroarylalkyl, alkoxy, alkoxyalkyl, or hydroxyalkyl is optionally substituted with one or more substituents —OH, halo, or alkoxy;   each R 5  is independently selected from the group consisting of aminoalkyl, alkylaminoalkyl, aminoalkylaminoakyl, guanidinoalkyl, N-heterocyclylalkyl, N-heteroaryl, hydroxyalkyl, hydroxyether, alkoxyalkyl, and hydroxylheteroalkyl;   R 7  is NH 2 ; and   each of R a  and R b  is H.   
     
     
         45 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 43 , wherein:
 each R 3  independently is   
       
         
           
           
               
               
           
         
         each R 4  independently is selected from the group consisting of 
       
       
         
           
           
               
               
           
         
       
       and
 each R 5  is independently selected from the group consisting of 
 
       
         
           
           
               
               
           
         
       
     
     
         46 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 1 , wherein m is 0; n is 0; s is 3 or 4; t is 0; r is 3; o is 0; q is 2; and p is 1, and having a structure: 
       
         
           
           
               
               
           
         
       
     
     
         47 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 46 , wherein:
 R 1  is H or alkyl;   each R 3  independently is C 8 -C 24 alkyl;   each R 4  is independently a neutral spacer moiety that is a C 1 -C 4 -alkyl substituted by cycloalkyl, heterocyclylalkyl, alkylaryl, arylalkyl, alkylheteroaryl, heteroarylalkyl, alkoxy, alkoxyalkyl, or hydroxyalkyl, wherein each cycloalkyl, heterocyclylalkyl, alkylaryl, arylalkyl, alkylheteroaryl, heteroarylalkyl, alkoxy, alkoxyalkyl, or hydroxyalkyl is optionally substituted with one or more substituents —OH, halo, or alkoxy;   each R 5  is independently selected from the group consisting of aminoalkyl, alkylaminoalkyl, aminoalkylaminoakyl, guanidinoalkyl, N-heterocyclylalkyl, and N-heteroaryl, hydroxyalkyl, hydroxyether, alkoxyalkyl, and hydroxylheteroalkyl;   R 7  is NH 2 ; and   each of R a  and R b  is H.   
     
     
         48 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 46 , wherein:
 each R 3  independently is   
       
         
           
           
               
               
           
         
         each R 4  independently is selected from the group consisting of 
       
       
         
           
           
               
               
           
         
       
       and
 each R 5  is independently selected from the group consisting of 
 
       
         
           
           
               
               
           
         
       
     
     
         49 . A tertiary amino lipidated and PEGylated peptide compound of formula (I) or salt thereof: 
       
         
           
           
               
               
           
         
         wherein: 
         m is an integer from 0 to 10; 
         n is an integer from 0 to 5; 
         s is an integer from 0 to 10; 
         t is an integer from 0 to 5;
 wherein at least one of m and t is nonzero, and wherein at least one of n and s is nonzero; 
 
         r is an integer from 1 to 20; 
         each o is independently an integer 0, 1, or 2; 
         each q is independently an integer 0, 1, or 2; 
         each p is independently an integer 1 or 2; 
         R 1  is —H, alkyl, alkylaryl, —COR 1a  or a lipid moiety, 
         wherein R 1a  is —H, —OH, alkyl, aryl, alkylaryl, —O-alkyl, or —O-alkylaryl; 
         each R 2  is independently an ethylene glycol moiety of the formula —CH 2 CH 2 O(CH 2 CH 2 O) u CH 3 , and wherein each u is independently an integer from 2 to 200; 
         each R 3  is independently a lipid moiety; 
         each R 4  is independently a neutral spacer moiety or a lipid moiety; 
         each R 5  is independently a cationic moiety; 
         each R 6  is independently an ethylene glycol moiety of the formula 
         CH 2 CH 2 O(CH 2 CH 2 O) v CH 3 , and wherein each v is independently an integer from 2 to 200; 
         R 7  is —H, alkyl, acyl, —OH, —OR 7a , —NH 2 , —NHR 7a , or a lipid moiety, wherein R 7a  is alkyl, acyl, or a lipid moiety; and 
         each R a  and R b  is independently —H, C 1 -C 4 -alkyl, or a side chain moiety found on a naturally- or non-naturally-occurring amino acid. 
       
     
     
         50 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 49 , wherein m is 1; n is 0; s is 3 or 4; t is 0; r is 1 or 2; o is 0; q is 2; and p is 1, and having a structure: 
       
         
           
           
               
               
           
         
       
     
     
         51 . The tertiary amino lipidated and/or PEGylated cationic peptide compound of  claim 50 , wherein:
 R 1  is H;   R 2  is an ethylene glycol moiety of the formula —CH 2 CH 2 O(CH 2 CH 2 O) u CH 3 , and wherein each u is independently an integer from 2 to 50;   each R 3  independently is C 8 -C 24 alkyl;   each R 4  is independently a neutral spacer moiety that is a C 1 -C 4 -alkyl substituted by cycloalkyl, heterocyclylalkyl, alkylaryl, arylalkyl, alkylheteroaryl, heteroarylalkyl, alkoxy, alkoxyalkyl, or hydroxyalkyl, wherein each cycloalkyl, heterocyclylalkyl, alkylaryl, arylalkyl, alkylheteroaryl, heteroarylalkyl, alkoxy, alkoxyalkyl, or hydroxyalkyl is optionally substituted with one or more substituents —OH, halo, or alkoxy;   each R 5  is independently selected from the group consisting of aminoalkyl, alkylaminoalkyl, aminoalkylaminoakyl, guanidinoalkyl, N-heterocyclylalkyl, and N-heteroaryl, hydroxyalkyl, hydroxyether, alkoxyalkyl, and hydroxylheteroalkyl;   R 7  is NH 2 ; and   each of R a  and R b  is H.   
     
     
         52 . A complex comprising one or more of the tertiary amino lipidated and/or PEGylated cationic peptide compounds of  claim 1 , complexed to a polyanionic compound. 
     
     
         53 . The complex of  claim 52 , wherein the polyanionic compound comprises a nucleic acid. 
     
     
         54 . The complex of  claim 52 , wherein the polyanionic compound comprises a nucleic acid and wherein the nucleic acid is an mRNA encoding a polypeptide. 
     
     
         55 . The complex of  claim 52 , wherein the polyanionic compound comprises a nucleic acid and wherein the complex comprises at least one of the tertiary amino lipidated and/or PEGylated cationic peptide and the nucleic acid at a mass ratio of between 0.5:1 and 50:1. 
     
     
         56 . The complex of  claim 52 , wherein the complex comprises: the one or more tertiary amino lipidated and/or PEGylated cationic peptide compounds, a nucleic acid, a phospholipid, a structural lipid, and a PEG lipid. 
     
     
         57 . A method of delivering a polyanionic compound to a cell comprising contacting the cell with the complex of  claim 52 . 
     
     
         58 . The method of  claim 57 , wherein the polyanionic compound comprises a nucleic acid which is the mRNA encoding a polypeptide and the cell expresses the polypeptide after being contacted with the complex. 
     
     
         59 . A method of forming the complex of  claim 52 , comprising contacting the tertiary amino lipidated and/or PEGylated cationic peptide compound with the polyanionic compound. 
     
     
         60 . A compound selected from the compounds listed in Table 1A.